ATF4 To Detect The Presence Of ER Stress

Food Science and Human Nutrition
Colorado State University
Graduate Student

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Company:

Abcam

Product Name:

Anti-ATF4 antibody

Catalog Number:

ab184909

Browse Similar Products: ATF4 Western Blot Antibodies

PERK is a key protein that is phosphorylated and activated during ER stress, responsible for the phosphorylation eIF2alpha which then activates ATF4. Determining the amount of ATF4 present due to a variety of stress conditions is helpful in assessing the severity of ER stress.

Experimental Design and Results Summary

Applications

Western Blot

Sample

Total lysates of H4IIE rat hepatoma cells

Primary Incubation

1:500 in 1:3 odyssey block:1X TBST (0.1% Tween) shaking, overnight at 4 degrees C

Blocking Agent

1:3 odyssey block:1X TBS at room temperature for 1 hr.

Secondary Incubation

1:15000 Goat anti-rabbit IRDye in 1:3 odyssey block:1X TBST (0.1% Tween) and 0.2% SDS for one hour at room temperature in the dark

Tertiary Incubation

N/A

Detection

Licor Odyssey Clx

Results Summary

This antibody works and appears around 38kDa, just like it is predicted. However, we attempted to use a blocking peptide for this specific antibody and that did not work. Therefore I am concerned this visible band may be non-specific binding.

DOI or PMID #

N/A

Additional Notes

N/A

Related Categories

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Summary

The Good

Shows up...

The Bad

May not be the correct protein

The Bottom Line

Order this antibody only if you can confirm with a blocking peptide this is the correct band.

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